Lipid formulations for nucleic acid delivery

Abstract

The present invention provides novel, stable lipid particles comprising one or more active agents or therapeutic agents, methods of making the lipid particles, and methods of delivering and/or administering the lipid particles. More particularly, the present invention provides stable nucleic acid-lipid particles (SNALP) comprising a nucleic acid (such as one or more interfering RNA), methods of making the SNALP, and methods of delivering and/or administering the SNALP.

1. A nucleic acid-lipid particle comprising:
 (a) a nucleic acid; 
 (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle; 
 (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof, wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle; and 
 (d) a conjugated lipid that inhibits aggregation of particles comprising from 0.5 mol % to 2 mol % of the total lipid present in the particle. 
 
     
     
       2. The nucleic acid-lipid particle of  claim 1 , wherein the nucleic acid comprises a small interfering RNA (siRNA). 
     
     
       3. The nucleic acid-lipid particle of  claim 2 , wherein the siRNA comprises from about 15 to about 60 nucleotides. 
     
     
       4. The nucleic acid-lipid particle of  claim 2 , wherein the siRNA comprises at least one modified nucleotide. 
     
     
       5. The nucleic acid-lipid particle of  claim 2 , wherein the siRNA comprises at least one 2′-O-methyl (2′OMe) nucleotide. 
     
     
       6. The nucleic acid-lipid particle of  claim 2 , wherein said siRNA is about 19 to about 25 base pairs in length. 
     
     
       7. The nucleic acid-lipid particle of  claim 2 , wherein said siRNA comprises 3′ overhangs. 
     
     
       8. The nucleic acid-lipid particle of  claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present in the particle. 
     
     
       9. The nucleic acid-lipid particle of  claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), or a mixture thereof. 
     
     
       10. The nucleic acid-lipid particle of  claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises a polyethyleneglycol (PEG)-lipid conjugate. 
     
     
       11. The nucleic acid-lipid particle of  claim 10 , wherein the PEG-lipid conjugate comprises a PEG-diacylglycerol (PEG-DAG) conjugate, a PEG-dialkyloxypropyl (PEG-DAA) conjugate, or a mixture thereof. 
     
     
       12. The nucleic acid-lipid particle of  claim 11 , wherein the PEG-DAA conjugate comprises a PEG-dimyristyloxypropyl (PEG-DMA) conjugate, a PEG-distearyloxypropyl (PEG-DSA) conjugate, or a mixture thereof. 
     
     
       13. The nucleic acid-lipid particle of  claim 12 , wherein the PEG has an average molecular weight of about 2,000 daltons. 
     
     
       14. The nucleic acid-lipid particle of  claim 10 , wherein the nucleic acid-lipid particle comprises about 57.1 mol % cationic lipid, about 7.1 mol % phospholipid, about 34.3 mol % cholesterol or a derivative thereof, and about 1.4 mol % PEG-lipid conjugate. 
     
     
       15. The nucleic acid-lipid particle of  claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present in the particle. 
     
     
       16. The nucleic acid-lipid particle of  claim 1 , wherein the nucleic acid in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C. for 30 minutes. 
     
     
       17. The nucleic acid-lipid particle of  claim 1 , wherein the nucleic acid is fully encapsulated in the nucleic acid-lipid particle. 
     
     
       18. The nucleic acid-lipid particle of  claim 1 , wherein the nucleic acid-lipid particle has a lipid:nucleic acid mass ratio of from about 5 to about 15. 
     
     
       19. The nucleic acid-lipid particle of  claim 1 , wherein the nucleic acid-lipid particle has a median diameter of from about 40 nm to about 150 nm. 
     
     
       20. The nucleic acid-lipid particle of  claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present in the particle. 
     
     
       21. The nucleic acid-lipid particle of  claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present in the particle. 
     
     
       22. A pharmaceutical composition comprising a nucleic acid-lipid particle of  claim 1  and a pharmaceutically acceptable carrier.